基于real-time PCR的齿垢密螺旋体含量与慢性牙周炎严重程度的关系研究

摘要

BACKGROUND:Oral treponemes (especially Treponema denticola) are widely considered to play important roles in the occurrence and development of chronic peridontitis.Treponema denticola can be detected in both periodontal patients and healthy individuals,so its pathogenicity may depend on its relative numbers in subgingival plaque.OBJECTIVE:To detect the number of Treponema denticola in subgingival plaque of patients with different severities of chronic periodontitis and to investigate the relationship between the bacterial colonization and the periodontal status.METHODS:Subgingival plaque samples were respectively taken from 132 patients with chronic periodontitis and 40 healthy controls,admitted in the Department of Stomatology,Aviation General Hospital of Chinese Medical University from July 2015 to August 2016.Based on the clinical data,132 patients were divided into mild (n=41),moderate (n=46),and severe (n=45) groups.The presence,relative numbers and proportion of Treponema denticola in the samples were detected by TaqMan real-time PCR assay.RESULTS AND CONCLUSION:There was no difference in the presence of Treponema dentico/a in subgingival plaque between healthy and periodontal participants.The relative numbers and proportion of Treponema denticola in the periodontal patients were significantly higher than those in the healthy controls (P ＜ 0.000 1).The quantification of Treponema denticola in different stages of periodontitis did show significant differences (P ＜ 0.05).In addition,the bacterial level and proportion of Treponema denticola were positively correlated to the probing depths (P ＜ 0.000 1).When the probing depth was more than 7 mm,the bacterial level and proportion of Treponema denticola were significantly increased (P ＜ 0.05,P ＜ 0.01).These findings suggest that Treponema denticola widely exists in subgingival plaque of human.The bacterial load and proportion of Treponema denticola are tightly related to the severity of periodontitis and probing depth.The higher bacterial level and proportion of Treponema denticola indicate a severer periodontitis.Moreover,real-time PCR possesses a broad potential in the studies on etiology,diagnosis and treatment of pedodontology.%背景:口腔密螺旋体与慢性牙周炎的发生发展密切相关.其中齿垢密螺旋体被认为是中等证据的致病菌,常在牙周炎患者龈下菌斑中检出,但在牙周健康人群中也能检测到,其致病性可能与在牙周的定植数量有关.目的:探讨齿垢密螺旋体与慢性牙周炎不同发展阶段以及相应的牙周临床指标的关系.方法:选择2015年7月至2016年8月在航空总医院口腔科收治的132例慢性牙周炎患者和40例牙周健康者,按照临床检查资料评估,将慢性牙周炎患者分为轻度组(n=41)、中度组(n=46)和重度组(n=45).收集所有患者的龈下菌斑,应用TaqMan实时荧光定量PCR技术,检测龈下菌斑中齿垢密螺旋体的检出率、相对检出量和所占比例.结果与结论:①齿垢密螺旋体在健康人(100％)和慢性牙周炎患者(100％)之间检出率差异无显著意义;②慢性牙周炎组齿垢密螺旋体的相对数量及所占比例均显著高于健康组(P＜0.000 1),轻、中和重度慢性牙周炎组之间两两比较差异均具有统计学意义(P＜ 0.05);③齿垢密螺旋体检出量和所占比例与牙周探诊深度呈显著正相关(P＜ 0.0001),当探诊深度≥7mm时,齿垢密螺旋体检出量明显升高(P＜0.05),齿垢密螺旋体所占比例也显著升高(P＜0.01);④结果表明,齿垢密螺旋体在健康人和慢性牙周炎患者的龈下菌斑中普遍存在.龈下菌斑中齿垢密螺旋体定植水平和所占比例与牙周炎严重程度、牙周探诊深度密切相关.定植数量越高,所占比例越多,则患者罹患牙周炎越严重.实时荧光定量PCR对牙周病病因、诊断及治疗研究具有应用前景.